PCCA Knockout cell line(Hep G2)

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PCCA Knockout cell line(Hep G2)

PCCA Knockout cell line(Hep G2) Order

Catalog#YKO-H514

Price (USD)-

Size 1*10^6

Instruction

Exclusive Red Cotton system(en.rc-crispr.com)was used to design the sgRNA fragment knockout strategy. Two sgRNAs were constructed into CRISPR-U™ plasmid which will be subsequentially transfected into cells by electroporation/virus method. And the single-cell clones were isolated by the limited dilution method and the Monoclone Genotype Validation Kit (Cat: YK-MV-1000) was used for cell lysis and PCR amplification on the single-cell clones, and then the genotype was verified by Sanger sequencing. Upon passing the final validation, the positive clones were expanded and cryopreserved.

Cell Info

Catalog YKO-H514
Cell line PCCA Knockout cell line(Hep G2) Cell Type Human Hepatoma Cell Line
Morphology Epithelial-like, adherent Passage ratio 1:3~1:5
Culture method 90% DMEM+10% FBS
Cryopreservation solution 50%DMEM+40%FBS+10%DMSO
Special Note

STR Validation

Gene Information

Official symbol
PCCA
Gene id
5095
Official full symbol
propionyl-CoA carboxylase subunit alpha
Also known as
None
The protein encoded by this gene is the alpha subunit of the heterodimeric mitochondrial enzyme Propionyl-CoA carboxylase. PCCA encodes the biotin-binding region of this enzyme. Mutations in either PCCA or PCCB (encoding the beta subunit) lead to an enzyme deficiency resulting in propionic acidemia. Multiple transcript variants encoding different isoforms have been found for this gene.

KO strategy

Knockout region: Exon 7 (based on transcript PCCA-202)
Validation: PCR+Sanger Sequencing

Method

Exclusive Red Cotton system(en.rc-crispr.com)was used to design the sgRNA fragment knockout strategy. Two sgRNAs were constructed into CRISPR-U™ plasmid which will be subsequentially transfected into cells by electroporation/virus method. And the single-cell clones were isolated by the limited dilution method and the Monoclone Genotype Validation Kit (Cat: YK-MV-1000) was used for cell lysis and PCR amplification on the single-cell clones, and then the genotype was verified by Sanger sequencing. Upon passing the final validation, the positive clones were expanded and cryopreserved.

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